丹参酮对乳腺癌4T1细胞增殖、CXCL1表达的影响及对Treg的调节作用  

Effect of tanshinone on the proliferation of 4T1 breast cancer cells,CXCL1 expression and its regulatory role on Treg cells

作　　者：刘才香 袁银胶 胡夏荣 罗锐娟 叶永康 黄林旋 郑锐年[1] LIU Caixiang;YUAN Yinjiao;HU Xiarong;LUO Ruijuan;YE Yongkang;HUANG Linxuan;ZHENG Ruinian(Department of Oncology,the 10th Affiliated Hospital of Southern Medical University/Dongguan People′s Hospital,Dongguan,Guangdong 523059,China;Department of General Surgery,the 10th Affiliated Hospital of Southern Medical University/Dongguan People′s Hospital,Dongguan,Guangdong 523059,China;Department of Breast Surgery,the 10th Affiliated Hospital of Southern Medical University/Dongguan People′s Hospital,Dongguan,Guangdong 523059,China;Department of Urology,the 10th Affiliated Hospital of Southern Medical University/Dongguan People′s Hospital,Dongguan,Guangdong 523059,China)

机构地区：[1]南方医科大学第十附属医院/广东省东莞市人民医院肿瘤科,广东东莞523059 [2]南方医科大学第十附属医院/广东省东莞市人民医院普外科,广东东莞523059 [3]南方医科大学第十附属医院/广东省东莞市人民医院乳腺科,广东东莞523059 [4]南方医科大学第十附属医院/广东省东莞市人民医院泌尿外科,广东东莞523059

出　　处：《检验医学与临床》2025年第4期522-525,530,共5页Laboratory Medicine and Clinic

基　　金：广东省基础与应用基础研究基金项目(2021A1515010156,2022A1515140134);广东省东莞市社会发展科技项目(20221800901582,20231800940482)。

摘　　要：目的探讨丹参酮对乳腺癌4T1细胞增殖、趋化因子配体1(CXCL1)表达的影响及对调节性T细胞(Treg)的调节作用。方法培养4T1细胞,设置对照组、5μg/mL丹参酮组和10μg/mL丹参酮组,对照组不进行处理,其余组加入相应浓度的药物处理24 h。采用细胞计数试剂盒(CCK-8)检测4T1细胞吸光度;采用定量聚合酶链反应及酶联免疫吸附试验(ELISA)分别检测4T1细胞中CXCL1 mRNA、蛋白的表达;采用流式细胞术检测原始CD4^(+)T细胞分化为Treg的情况。将10μg/mL丹参酮干预过的Treg,与CD8^(+)T细胞、4T1细胞按照1∶10∶2的比例共培养。采用ELISA试剂盒检测4T1细胞分泌的颗粒酶B、穿孔素水平。结果与对照组相比,5、10μg/mL丹参酮组吸光度,以及CXCL1 mRNA、蛋白表达水平明显降低,差异均有统计学意义(P<0.05)。与对照组相比,5μg/mL和10μg/mL丹参酮组原始CD4+T细胞分化为Treg的比例明显降低,差异均有统计学意义(P<0.05)。与对照组相比,5μg/mL丹参酮组和10μg/mL丹参酮组CD8^(+)T细胞分泌的颗粒酶B和穿孔素水平升高,差异均有统计学意义(P<0.05)。结论丹参酮可抑制乳腺癌4T1细胞的增殖和降低CXCL1 mRNA和蛋白的表达水平,抑制原始CD4^(+)T细胞分化为Treg。此外,丹参酮也可以抑制共培养体系中Treg对CD8^(+)T细胞的免疫抑制功能。Objective To investigate the effects of tanshinone on the proliferation of 4T1 breast cancer cells,the expression of chemokine ligand 1(CXCL1),and its regulatory effect on regulatory T cells(Treg).Methods 4T1 cells were cultured and divided into the control group,5μg/mL tanshinone group and 10μg/mL tanshinone group.The control group was not treated,while the other groups were treated with the corresponding concentrations of the drug for 24 h.The cell counting kit-8(CCK-8)was used to detect the absorbance of 4T1 cells.Quantitative polymerase chain reaction(qPCR)and enzyme-linked immunosorbent assay(ELISA)were used to detect the expression of CXCL1 mRNA and protein secreted by 4T1 cells.Flow cytometry was used to assess the differentiation of naive CD4^(+)T cells into Treg.The 10μg/mL tanshinone-treated Treg were co-cultured with CD8^(+)T cells and 4T1 cells at a ratio of 1∶10∶2.The levels of granzyme B and perforin secreted by CD8^(+)T cells were detected using an ELISA kit.Results Compared with the control group,the absorbance and the expression levels of CXCL1 mRNA and protein in the 5μg/mL tanshinone group and the 10μg/mL tanshinone group decreased significantly,the differences were statistically significant(P<0.05).Additionally,the proportion of naive CD4^(+)T cells differentiating into Treg was significantly lower in the 5μg/mL tanshinone group and the 10μg/mL tanshinone group compared with the control group,and the differences were statistically significant(P<0.05).Compared with the control group,the levels of granzyme B and perforin secreted by CD8^(+)T cells were higher in the 5μg/mL tanshinone group and the 10μg/mL tanshinone group,and the differences were statistically significant(P<0.05).Conclusion Tanshinone can inhibit the proliferation of breast cancer 4T1 cell and reduce the expression levels of CXCL1 mRNA and protein.It suppresses the differentiation of naive CD4^(+)T cells into Treg.Furthermore,tanshinone can inhibit the immunosuppressive function of Treg on CD8^(+)T cells in the co-c

关 键 词：丹参酮 乳腺癌 趋化因子配体 CD4^(+)T细胞 调节性T细胞 

分 类 号：R285.5[医药卫生—中药学] R737.9[医药卫生—中医学]